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Elife. 2019 May 24;8. pii: e44635. doi: 10.7554/eLife.44635.

Comprehensive substrate specificity profiling of the human Nek kinome reveals unexpected signaling outputs.

Author information

1
Department of Biology, Massachusetts Institute of Technology, Cambridge, United States.
2
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, United States.
3
Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, United States.
4
MIT Center for Precision Cancer Medicine, Massachusetts Institute of Technology, Cambridge, United States.
5
Department of Pharmacology, Yale School of Medicine, New Haven, United States.
6
Department of Medical Biophysics, University of Toronto, Toronto, Canada.
7
Biotech Research and Innovation Center, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
8
Princess Margaret Cancer Center, University Health Network, Toronto, Canada.
9
Department of Surgery, Beth Israel Deaconess Medical Center, Divisions of Acute Care Surgery, Trauma, and Critical Care and Surgical Oncology, Harvard Medical School, Boston, United States.

Abstract

Human NimA-related kinases (Neks) have multiple mitotic and non-mitotic functions, but few substrates are known. We systematically determined the phosphorylation-site motifs for the entire Nek kinase family, except for Nek11. While all Nek kinases strongly select for hydrophobic residues in the -3 position, the family separates into four distinct groups based on specificity for a serine versus threonine phospho-acceptor, and preference for basic or acidic residues in other positions. Unlike Nek1-Nek9, Nek10 is a dual-specificity kinase that efficiently phosphorylates itself and peptide substrates on serine and tyrosine, and its activity is enhanced by tyrosine auto-phosphorylation. Nek10 dual-specificity depends on residues in the HRD+2 and APE-4 positions that are uncommon in either serine/threonine or tyrosine kinases. Finally, we show that the phosphorylation-site motifs for the mitotic kinases Nek6, Nek7 and Nek9 are essentially identical to that of their upstream activator Plk1, suggesting that Nek6/7/9 function as phospho-motif amplifiers of Plk1 signaling.

KEYWORDS:

Nek kinase family; Nima-related kinase; biochemistry; cancer biology; chemical biology; kinase biology; mitosis; none; signaling; substrate specificity

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